<p>Trypanosomes were purified from infected blood, fixed and processed for immunofluorescence using the affinity purified α-FLA3 antibodies. Upper panels present a merge of the fluorescence and phase images and reveal the location of FLA3 (green) and the nucleus/kinetoplast (blue); lower panels present the fluorescence image alone. Bar = 10 µm. Panel A. FLA3 was detected as a punctate pattern of staining along the FAZ. The black arrows in the upper panel (α-FLA3) indicate the absence of FLA3 along the free flagellum. Non specific binding was investigated by including the peptide antigen (10 µg.ml<sup>−1</sup>) (α-FLA3+ pep). Panel B. FLA3 is associated with existing and new (arrows) FAZ. Panel C. FLA3 remains associated with the cell body at regions where the flagellum has become detached (black arrows in the phase/fluorescence merge). The white arrows indicate more pronounced staining of FLA3 frequently observed at the posterior and anterior end of the attachment zone (lower panels).</p
